The DOI matches its Crossref record; 26 of the 26 references it cites were found in a registry or an index.
Amplicon DNA Melting Analysis for Mutation Scanning and Genotyping: Cross-Platform Comparison of Instruments and Dyes
Abstract
Background: DNA melting analysis for genotyping and mutation scanning of PCR products by use of high-resolution instruments with special “saturation” dyes has recently been reported. The comparative performance of other instruments and dyes has not been evaluated. Methods: A 110-bp fragment of the β-globin gene including the sickle cell anemia locus (A17T) was amplified by PCR in the presence of either the saturating DNA dye, LCGreen Plus, or SYBR Green I. Amplicons of 3 different genotypes (wild-type, heterozygous, and homozygous mutants) were melted on 9 different instruments (ABI 7000 and 7900HT, Bio-Rad iCycler, Cepheid SmartCycler, Corbett Rotor-Gene 3000, Idaho Technology HR-1 and LightScanner, and the Roche LightCycler 1.2 and LightCycler 2.0) at a rate of 0.1 °C/s or as recommended by the manufacturer. The ability of each instrument/dye combination to genotype by melting temperature (Tm) and to scan for heterozygotes by curve shape was evaluated. Results: Resolution varied greatly among instruments with a 15-fold difference in Tm SD (0.018 to 0.274 °C) and a 19-fold (LCGreen Plus) or 33-fold (SYBR Green I) difference in the signal-to-noise ratio. These factors limit the ability of most instruments to accurately genotype single-nucleotide polymorphisms by amplicon melting. Plate instruments (96-well) showed the greatest variance with spatial differences across the plates. Either SYBR Green I or LCGreen Plus could be used for genotyping by Tm, but only LCGreen Plus was useful for heterozygote scanning. However, LCGreen Plus could not be used on instruments with an argon laser because of spectral mismatch. All instruments compatible with LCGreen Plus were able to detect heterozygotes by altered melting curve shape. However, instruments specifically designed for high-resolution melting displayed the least variation, suggesting better scanning sensitivity and specificity. Conclusion: Different instruments and dyes vary widely in their ability to genotype homozygous variants and scan for heterozygotes by whole-amplicon melting analysis.
Overview
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Integrity signals
Registry
Cites
Statistics
Verification
Claims the paper makes
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
The self-check needs the paper’s full text.
Citations
Cites
Saiki, R. K., Scharf, S., Faloona, F., Mullis, K. B., Horn, G. T., Erlich, H. A., & Arnheim, N. (1985) · Science · doi:10.1126/science.2999980 · Cited by 8,956
A unified view of polymer, dumbbell, and oligonucleotide DNA nearest-neighbor thermodynamics
SantaLucia, J. (1998) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.95.4.1460 · Cited by 2,793
Product Differentiation by Analysis of DNA Melting Curves during the Polymerase Chain Reaction
Ririe, K. M., Rasmussen, R. P., & Wittwer, C. T. (1997) · Analytical Biochemistry · doi:10.1006/abio.1996.9916 · Cited by 1,654
Self-citationContinuous Fluorescence Monitoring of Rapid Cycle DNA Amplification
Wittwer, C. T., Herrmann, M. G., Moss, A. A., & Rasmussen, R. P. (2013) · BioTechniques · doi:10.2144/000114043 · Cited by 1,274
Self-citationHigh-Resolution Genotyping by Amplicon Melting Analysis Using LCGreen
Wittwer, C. T., Reed, G. H., Gundry, C. N., Vandersteen, J. G., & Pryor, R. J. (2003) · Clinical Chemistry · doi:10.1373/49.6.853 · Cited by 1,257
Self-citationContinuous Fluorescence Monitoring of Rapid Cycle DNA Amplification
Wittwer, C. T., Herrmann, M. G., Moss, A. A., & Rasmussen, R. P. (1997) · BioTechniques · doi:10.2144/97221bi01 · Cited by 1,143
Self-citationQuantification of low-copy transcripts by continuous SYBR Green I monitoring during amplification.
Morrison, T. B., Weis, J. J., & Wittwer, C. T. (1998) · PubMed · Cited by 958
Self-citationThe LightCycler TM : A Microvolume Multisample Fluorimeter with Rapid Temperature Control
Wittwer, C. T., Ririe, K. M., Andrew, R. V., David, D. A., Gundry, R. A., & Balis, U. J. (1997) · BioTechniques · doi:10.2144/97221pf02 · Cited by 893
Self-citationZipper, H. (2004) · Nucleic Acids Research · doi:10.1093/nar/gnh101 · Cited by 791
Genotyping of Single-Nucleotide Polymorphisms by High-Resolution Melting of Small Amplicons
Liew, M., Pryor, R., Palais, R., Meadows, C., Erali, M., Lyon, E., & Wittwer, C. (2004) · Clinical Chemistry · doi:10.1373/clinchem.2004.032136 · Cited by 660
Self-citation
Cited by
High-Resolution DNA Melting Analysis for Simple and Efficient Molecular Diagnostics
Reed, G. H., Kent, J. O., & Wittwer, C. T. (2007) · Pharmacogenomics · doi:10.2217/14622416.8.6.597 · Cited by 678
Self-citationHigh-Resolution Melting Analysis (HRMA)-More than just sequence variant screening
Vossen, R. H. A. M., Aten, E., Roos, A., & den Dunnen, J. T. (2009) · Human Mutation · doi:10.1002/humu.21019 · Cited by 484
High-resolution DNA melting analysis: advancements and limitations
Wittwer, C. T. (2009) · Human Mutation · doi:10.1002/humu.20951 · Cited by 468
Self-citationAn update on current and novel molecular diagnostics for the diagnosis of invasive fungal infections
Jenks, J. D., White, P. L., Kidd, S. E., Goshia, T., Fraley, S. I., Hoenigl, M., & Thompson, G. R. (2023) · Expert Review of Molecular Diagnostics · doi:10.1080/14737159.2023.2267977 · Cited by 61
Metadata
- Authors
- Herrmann, M. G., Durtschi, J. D., Bromley, L. K., Wittwer, C. T., & Voelkerding, K. V.
- Venue
- Clinical Chemistry
- Publisher
- Oxford University Press (OUP)
- Published
- 2006-03-01
- Volume
- 52
- Issue
- 3
- Pages
- 494-503
- ISSN
- 0009-9147 (ISSN-L)
- 1530-8561
- OpenAlex
- W2063542361
- Cited by
- 279
- References
- 26
- FWCI
- 12.16
- Type
- journal article
- Language
- en
- Open access
- closed
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
How to cite
Cite CoChat for the checks on this page, and the paper for its own findings.
- This page
- CoChat. (2026). Citation check: Amplicon DNA Melting Analysis for Mutation Scanning and Genotyping: Cross-Platform Comparison of Instruments and Dyes. https://app.cochat.ai/papers/10.1373/clinchem.2005.063438
- The paper
- Herrmann, M. G., Durtschi, J. D., Bromley, L. K., Wittwer, C. T., & Voelkerding, K. V. (2006). Amplicon DNA Melting Analysis for Mutation Scanning and Genotyping: Cross-Platform Comparison of Instruments and Dyes. Clinical Chemistry. https://doi.org/10.1373/clinchem.2005.063438
Data sources
Bibliographic data: Semantic Scholar, OpenAlex, Crossref.
Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).
Semantic Scholar’s data is described in: Kinney, R., et al. (2023). The Semantic Scholar Open Data Platform. arXiv:2301.10140.
